CAJ | 학술논문

目的探讨诱导痰处理温度对哮喘患者诱导痰中炎性细胞分类及其气道炎性反应标志物浓度的影响.方法收集2013年3月至2013年8月在广州呼吸疾病研究所就诊的哮喘患者52例,所有受试者进行痰诱导,选取痰黏液加入4倍体积0.1％的二硫苏糖醇(DTT)后,分别于0℃冰浴与37℃水浴下孵育15 min(15 min冰浴组和15 min水浴组,n=28)和30 min(30 min冰浴组和30 min水浴组,n=10).细胞沉淀行HE染色后进行分类计数,同时采用悬浮芯片方法检测痰上清白细胞介素(IL)-5、IL-10、IL-17、干扰素(IFN)-γ和肿瘤坏死因子(TNF)-α.比较不同诱导痰处理温度对痰细胞计数及气道炎性反应指标水平的影响.结果 52例哮喘患者,痰诱导成功28例,成功率为53.8％.15 min冰浴组和15 min水浴组比较、30 min冰浴组和30 min水浴组比较,其痰液重量、痰细胞总数、细胞存活率和痰细胞分类差异均无统计学意义(均P＞0.05).15 min冰浴组和15 min水浴组痰上清IL-10、IL-17、IFN-γ、IL-5和TNF-α的浓度比较差异无统计学意义(均P＞0.05);30 min冰浴组和30 min水浴组痰上清IL-10、IL-17和IFN-γ的浓度差异无统计学意义(均P＞0.05),但30 min水浴组的IL-5和TNF-α浓度要高于30min冰浴组[IL-5:2.55 (2.20～4.35) ng/L比2.03(1.52～2.50) ng/L,TNF-α:19.41(9.07～37.92) ng/L比10.78(8.58～ 17.93)ng/L,均P＜0.05].结论诱导痰处理温度会影响诱导痰中某些气道炎性反应标志物的浓度.在诱导痰处理时DTT裂解后,置于0℃冰浴下操作,可以较真实反映诱导痰中炎性反应标志物的真实浓度.
목적 탐토유도담처리온도대효천환자유도담중염성세포분류급기기도염성반응표지물농도적영향.방법 수집2013년3월지2013년8월재엄주호흡질병연구소취진적효천환자52례,소유수시자진행담유도,선취담점액가입4배체적0.1％적이류소당순(DTT)후,분별우0℃빙욕여37℃수욕하부육15 min(15 min빙욕조화15 min수욕조,n=28)화30 min(30 min빙욕조화30 min수욕조,n=10).세포침정행HE염색후진행분류계수,동시채용현부심편방법검측담상청백세포개소(IL)-5、IL-10、IL-17、간우소(IFN)-γ화종류배사인자(TNF)-α.비교불동유도담처리온도대담세포계수급기도염성반응지표수평적영향.결과 52례효천환자,담유도성공28례,성공솔위53.8％.15 min빙욕조화15 min수욕조비교、30 min빙욕조화30 min수욕조비교,기담액중량、담세포총수、세포존활솔화담세포분류차이균무통계학의의(균P＞0.05).15 min빙욕조화15 min수욕조담상청IL-10、IL-17、IFN-γ、IL-5화TNF-α적농도비교차이무통계학의의(균P＞0.05);30 min빙욕조화30 min수욕조담상청IL-10、IL-17화IFN-γ적농도차이무통계학의의(균P＞0.05),단30 min수욕조적IL-5화TNF-α농도요고우30min빙욕조[IL-5:2.55 (2.20～4.35) ng/L비2.03(1.52～2.50) ng/L,TNF-α:19.41(9.07～37.92) ng/L비10.78(8.58～ 17.93)ng/L,균P＜0.05].결론 유도담처리온도회영향유도담중모사기도염성반응표지물적농도.재유도담처리시DTT렬해후,치우0℃빙욕하조작,가이교진실반영유도담중염성반응표지물적진실농도.
Objective To evaluate the effects of temperature on cytology of inflammatory cell and detection of inflammatory markers during sputum processing.Methods Fifty-two asthmatic patients were recruited from Guangzhou Institute of Respiratory Diseases between March and August 2013.Following sputum induction,0.1％ dithiothreitol (DTT),at a ratio of 4∶1 (v/w),was added for incubation consecutively at 0 ℃ and 37 ℃,for 15 min (15 min ice-incubated group and 15 min water bath group,n=28) and 30 min (30 min ice-incubated group and 30 min water bath group,n=10).The cell pellet was re-suspended for cytology assessment followed by hematoxylin-eosin (HE) staining.Commercially available luminex kits were employed to measure the levels of interleukin (IL)-5,IL-10,IL-17,interferon (IFN)-γ and tumor necrosis factor (TNF)-α in the sputum supernatant.Effects of different temperatures on cytology of inflammatory cell and detection of inflammatory markers during sputum processing were subsequently compared.Results Sputum induction was successful in 28 out of 52 patients (success rate 53.8％).There was no difference in sputum weight,total cell counts,cell viability and differential cell counts between the 15 min ice-incubated group and water bath group,30 min ice-incubated group and water bath group (all P＞0.05).There were no differences in the levels of IL-5,IL-10,IL-17,IFN-γ and TNF-α between 15 min ice-incubated group and 15 min water bath group (all P＞0.05).There were no differences in the levels of IL-10,IL-17 and IFN-γ between 30 min ice-incubated group and 30 min water bath group (all 9＞0.05).However,the levels of IL-5 and TNF-α in 30 min water bath group were significantly higher than those in 30 min ice-incubated group [2.55 (2.20-4.35) ng/L vs 2.03 (1.52-2.50) ng/L,19.41 (9.07-37.92) ng/L vs 10.78 (8.58-17.93) ng/L,all P＜0.05].Conclusions Levels of inflammatory mediators can be influenced by the different temperatures upon processing.Sputum processed with DTT at 0 ℃ may better reflect the true levels of inflammatory markers.